Characterisation and determination of metabolites formed by microbial and enzymatic degradation of ergot alkaloids

Abstract

Ergot alkaloids are frequent contaminants of cereal crops. Strategies for their inactivation include the use of microorganisms or enzymes as feed additives capable of degrading ergot alkaloids. Recently, an ergopeptine-degrading Rhodococcus erythropolis strain MTHt3 (DSM 25948) has been isolated from soil and the involved enzymes ErgA and ErgB have been identified. The aim of the current study was to characterise the metabolites formed by degradation of various ergopeptines with the MTHt3 strain, its lysate and the purified enzyme ErgA. Using preparative HPLC, 1H-, 13C- and 2D-NMR as well as HR-MS measurements, two main groups of metabolites formed during microbial and enzymatic degradation of ergopeptines were identified: diketopiperazines (cyclic dipeptides, DKPs) and unstable ergine hydroxy carboxylic acids. However, degradation by strain, lysate and enzyme yielded different end-products. Whereas DKPs were transient and lysergic acid the only final product upon incubation of ergopeptines with the R. erythropolis strain, incubation with the lysate resulted in formation of lysergic acid and two isomeric DKPs at different ratios. Enzymatic degradation by ErgA yielded only one DKP isomer and ended at the stage of the ergine hydroxy carboxylic acids which then spontaneously degraded to ergine. In conclusion, we succeeded in identification of metabolites formed by microbial and enzymatic degradation of ergot alkaloids which is a crucial step in the future development of feed additives for gastro intestinal detoxification of ergopeptines in farm animals.